乙型脑炎病毒感染猪睾丸细胞的miRNA表达谱差异分析

Identification and analysis of the miRNA expression profiles of Japanese encephalitis virus(JEV)infected swine testis(ST)cells

[目的]本试验旨在分析乙型脑炎病毒(JEV)感染猪睾丸(ST)细胞和正常ST细胞miRNA差异表达谱,以探索miRNA在JEV感染过程中的作用及其调控机制。[方法]分别提取正常ST细胞和JEV感染8h后的ST细胞总RNA,利用高通量测序技术检测分析ST细胞中的miRNA表达谱并进行差异分析。使用miRanda软件对表达差异显著miRNA的靶基因进行预测并对靶基因进行GO分析。选取6个显著性差异表达的miRNA通过RT-qPCR进行验证。[结果]与正常ST细胞相比,JEV感染后的ST细胞有112个差异表达miRNA,其中80个miRNA上调表达,32个miRNA下调表达。经RT-qPCR方法验证6个差异表达的miRNA结果与高通量测序结果一致。差异表达miRNA靶基因的生物学功能相对集中在细胞代谢、细胞黏附等生物过程。[结论]检测和分析JEV感染后猪睾丸细胞的miRNA表达谱,获得了大量的与JEV感染相关的miRNA表达谱数据,为揭示JEV感染生殖系统致病机制提供了有效的数据和理论基础。

[Objectives] The paper aimed to detect microRNA(miRNA)expression profile in Japanese encephalitis virus(JEV)infected swine testis(ST)cells.[Methods] Total RNA of JEV-infected and uninfected ST cells was extracted individually by Trizol and then analyzed miRNA expression profiles by high-throughput sequencing technology. miRNA target prediction algorithm miRanda was employed to predict potential target genes of differently expressed miRNA. Gene ontology(GO)analysis was performed on the host target genes,and 6 signifacant differences in expression of miRNA were verified by quantitative real time polymerase chain reaction( RT-qPCR ).[Results] The results indicated that total 112 miRNA were obviously differently expressed in the JEV-infected cells including 80 up-regulated miRNA and 32 down-regulated miRNA. The six of those miRNA were verified by RT-qPCR,the results were well matched with those of sequencing data. GO analysis of the target genes showed that these miRNA involved in the biological processes of cell metabolism and cell adhesion.[Conclusions] Our findings indicate that miRNA may play critical roles in the pathogenesis of JEV infection.